#8343 Jak/Stat Pathway Inhibitors Antibody Sampler Kit
|SOCS1 (A156) Antibody #3950||20 µl||WB||H, M, R, Mk||Dg||23||Rabbit|
|SOCS2 Antibody #2779||20 µl||WB, IP||H, M, R||Mk||22||Rabbit|
|SOCS3 (D6E1T) Rabbit mAb #52113||20 µl||WB||H, M||28||Rabbit IgG|
|PIAS1 (D33A7) XP® Rabbit mAb #3550||20 µl||WB, IF-IC, F||H, M, R, Mk||76||Rabbit IgG|
|PIAS3 (D5F9) XP® Rabbit mAb #9042||20 µl||WB, IP, IF-IC||H||Mk||65-75||Rabbit IgG|
|PIAS4 (D2F12) Rabbit mAb #4392||20 µl||WB||H, R, Mk||M||75||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||WB||Goat|
†Species predicted to react based on 100% sequence homology.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from K562, A204 and PC12 cells using SOCS2 Antibody.
Western blot analysis of extracts from various cell lines using PIAS1 (D33A7) XP® Rabbit mAb.
Western blot analysis of extracts from 293 and CCRF-CEM cells using PIAS4 (D2F12) Rabbit mAb.
Western blot analysis of extracts from BaF3 cells starved of mouse IL-3 and serum overnight, untreated (-) or mouse IL-3-treated (10 ng/ml, 6 hr; +) and NK-92 cells starved of human IL-2 and serum overnight, untreated (-) or human IL-2-treated (10 ng/ml, 6 hr; +), using SOCS1 (A156) Antibody.
Western blot analysis of extracts from RD cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® PIAS3 siRNA I #9073 (+), or SignalSilence® PIAS3 siRNA II #9031 (+), using PIAS3 (D5F9) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The PIAS3 (D5F9) XP® Rabbit mAb confirms silencing of PIAS3 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human SOCS3 (+), using SOCS3 (D6E1T) Rabbit mAb.
Flow cytometric analysis of HeLa cells using PIAS1 (D33A7) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Western blot analysis of extracts from COS-7 cells, untransfected (-) or transfected with a construct overexpressing human SOCS1 (+), using SOCS1 (A156) Antibody.
Western blot analysis of extracts from various cell lines using PIAS3 (D5F9) XP® Rabbit mAb.
Western blot analysis of extracts from Hep G2 and A-375 cell lines, untreated (-) or treated with Human Oncostatin M #5367 (hOSM; 20 ng/ml, 1 hr; +), using SOCS3 (D6E1T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Confocal imunofluorescent analysis of HeLa cells using PIAS1 (D33A7) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human PIAS3 (hPIAS3; +), using PIAS3 (D5F9) XP® Rabbit mAb.
Western blot analysis of extracts from KARPAS-299 cells, untreated (-) or treated with the Jak3 inhibitor WHI-P154 (40 μM, overnight; +), using SOCS3 (D6E1T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb (lower). KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.
Confocal immunofluorescent analysis of A-204 (positive, left) and Hep G2 (low expression, right) cells using PIAS3 (D5F9) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Jak (Janus Kinase) and Stat (signal transducer and activator of transcription) proteins are utilized by receptors for a wide varity of ligands including cytokines, hormones, growth factors, and neurotransmitters (1). Jaks and Stats play important roles in oncogenesis, tumor progression, angiogenesis, cell motility, immune responses, and stem cell differentiation (2-5). Therefore, regulation of Jak/Stat signaling is crucial to prevent aberrant signaling which can lead to disease progression. Two methods for regulating Jak/Stat signaling involve SOCS and PIAS proteins (6,7).
The SOCS (suppressor or cytokine signaling) family members are negative regulators of cytokine signal transduction that inhibit the Jak/Stat pathway and consist of 8 known members, including the originally identified protein CIS1 (cytokine-inducible SH2-containing protein) and SOCS1-SOCS7. Each SOCS family member contains a central SH2 domain and a conserved carboxy-terminal motif designated as the SOCS box. These proteins are important regulators of cytokine signaling, proliferation, differentiation, and immune responses (8-10). SOCS proteins are involved in regulating over 30 cytokines, including interleukins, growth hormone (GH), interferons, leptin, and leukemia inhibitory factor (7). SOCS1, also known as JAB (Janus Kinase binding protein) and SSI-1 (Stat-induced Stat inhibitor-1), shares the most homology with SOCS3 and both are highly induced by cytokines (7,11). Both SOCS1 and SOCS3 directly inhibit Jak activity; SOCS1 inhibits Jak through an interaction involving a phospohotyrosine located in the kinase activation loop; SOCS3 inhibits Jak via its SH2 domain (12,13). In addition to inhibiting Jak/Stat signaling, the SOCS box of SOCS1 and SOCS3 can trigger ubiquitin-mediated degradation of proteins within and outside the Jak/Stat pathway (14,15). SOCS2 is also incduced upon cytokine stimulation and the activity of SOCS2 has been predominately linked to GH and insulin-like growth factor signaling by binding to tyrosine-phosphorylated receptors via its SH2 domain (11,16).
The PIAS (protein inhibitor of activated Stats) proteins, which include PIAS1, PIAS3, PIASx, and PIASy (PIAS4), were originally characterized based on their interaction with the Stat family of transcription factors (16,17). PIAS1, PIAS3, and PIASx interact with and repress Stat1, Stat3, and Stat4, respectively (17-19). The PIAS family contains a conserved RING domain that has been linked to function as a SUMO (small ubiquitin-related modifer) ligase, coupling the SUMO conjugating enzyme Ubc9 with its substrate proteins leading to regulation of transcription factors through distinct mechanisms including NF-κB, c-Jun, Oct-4, p53, and SMADs. PIAS4 is a specific SUMO-E3 ligase for Ets-1 and represses Ets-1 dependent transcription in addition to altering the nuclear localization and reducing the transcriptional activity of C/EBPδ, thereby enhancing cell proliferation and migration (20,21).
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Tween is a registered trademark of ICI Americas, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
KARPAS cell line source: Dr. Abraham Karpas at the University of Cambridge.